Application of 3-methyladenine in preparation of medicine for preventing acute pneumonia induced by pseudomonas aeruginosa

A technology of Pseudomonas aeruginosa and methyladenine, which is applied in the field of biomedicine to achieve the effect of reducing inflammatory cytokines and bacterial load and avoiding the increasing drug resistance

Inactive Publication Date: 2021-07-20
INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although there are few in vivo reports on the role of autophagy in Pseudomonas aeruginosa infection, currently available in vivo reports on autophagy consistently show its negative role in infection and inflammation, including burns, mechanical ventilation, Porphyromonas gingivalis-induced periodontitis, lipopolysaccharide (LPS)-induced endothelial barrier dysfunction in acute lung injury, LPS-induced endotoxemia and polymicrobial sepsis, even methicillin-resistant Staphylococcus aureus pneumonia

Method used

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  • Application of 3-methyladenine in preparation of medicine for preventing acute pneumonia induced by pseudomonas aeruginosa
  • Application of 3-methyladenine in preparation of medicine for preventing acute pneumonia induced by pseudomonas aeruginosa
  • Application of 3-methyladenine in preparation of medicine for preventing acute pneumonia induced by pseudomonas aeruginosa

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Experimental program
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Effect test

Embodiment 1

[0027] Embodiment 1 - the preparation of bacterial liquid

[0028] The multidrug-resistant Pseudomonas aeruginosa clinical isolate 1409 was obtained from the First Affiliated Hospital of Kunming Medical University. Pseudomonas aeruginosa strain 8821 was a gift from A. Chakrabarty, University of Illinois at Chicago. Bacteria were grown with shaking in LB broth at 37°C and harvested when the culture reached an optical density of 2.5-3 at 600 nm (early stationary phase). Bacteria were washed with sterile phosphate-buffered saline (PBS) and then resuspended in saline for in vivo experiments or in PBS for in vitro experiments.

Embodiment 2

[0029] Embodiment 2——animal survival experiment

[0030] C57BL / 6 mice (female, 7-8 week-old specific pathogen-free C57BL / 6 mice were purchased from Life River Experimental Animal Technology Co., Ltd. (Beijing), and bred in the Experimental Animal Center of the Institute of Medical Biology, Chinese Academy of Medical Sciences) After tribromoethanol anesthesia, nasal infection 2×10 7 1409 strains of Pseudomonas aeruginosa with a colony forming unit (CFU) of 20 μL were isolated and monitored for 7 days. Disease scores were recorded as follows:

[0031] (1) Appearance (0-normal; 1-lack of grooming; 2-rough; 3-very coarse body hair);

[0032](2) Posture (0-normal; 1-sitting bowed; 2-bending posture, head resting on the floor; 3-prone at the bottom of the cage / unable to maintain an upright posture);

[0033] (3) Activity / behavior (0 - normal; 1 - slightly reduced / slightly altered behavior; 2 - above change plus changes in breathing rate or energy; 3 - only move when stimulated); ...

Embodiment 3

[0047] Embodiment 3——Pseudomonas aeruginosa pulmonary infection experiment

[0048] After anesthesia, mice were treated with 1×10 9 CFU nasal drops for infection. The volume of Pseudomonas aeruginosa 8821 was 20 μL. After 24 hours, inject 10 mL of PBS into the heart to remove blood from the lungs. The lungs were lavaged with 1 mL of PBS containing soybean trypsin inhibitor (100 μg / mL) to obtain bronchoalveolar lavage fluid (BALF), and then the alveoli were washed twice with 1 mL of PBS. BALF and lung tissues after lavage were collected, bacterial colony-forming units (CFU), macrophages and neutrophils were counted, and cytokine detection, MPO detection and histological examination were performed.

[0049] BALF (10 μL) was inoculated into LB solid culture dishes, and incubated for 24 hours to count CFU. To detect cytokines and MPO activity, BALF were centrifuged at 1200×g for 5 min at 4°C, and the supernatant was used for cytokine analysis. The pellet was resuspended in 1 ...

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Abstract

The invention relates to application of 3-methyladenine in preparation of a medicine for preventing acute pneumonia induced by pseudomonas aeruginosa, and belongs to the technical field of biomedicine. Experimental research shows that before pseudomonas aeruginosa infection, the survival rate of experimental animals can be increased by using 3-methyladenine, and meanwhile, host tissue damage caused by excessive inflammatory immune response is avoided along with pathology reduction and reduction of inflammatory cytokines and in-lung bacteria load; and the increasing drug resistance of bacteria to existing antibiotics due to improper use of the antibiotics can be avoided, and the application prospect is wide.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to the application of 3-methyladenine in the preparation of drugs for preventing acute pneumonia induced by Pseudomonas aeruginosa. Background technique [0002] Pseudomonas aeruginosa is one of the leading causes of nosocomial infections worldwide. Intensive care unit infections, especially ventilator-associated pneumonia, are associated with high mortality. In addition to high virulence, the tendency to develop resistance to multiple antimicrobial agents makes Pseudomonas aeruginosa difficult to control. The fact that clinical isolates of P. aeruginosa resistant to colistin and tigecycline have emerged has prompted us to look for solutions beyond traditional antibiotic therapy. In addition to immunization strategies that include vaccines (active immunization) and antibody administration (passive immunization), other host-targeted interventions have shown potentia...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/52A61P11/00A61P31/04
CPCA61K31/52A61P11/00A61P31/04
Inventor 刘存宝岳磊曹晗齐家龙王云飞李华
Owner INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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